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Effect of acute infection with noncytopathic or cytopathic bovine viral diarrhea virus isolates on bovine platelets
1994
Bezek, D.M. | Grohn, Y.T. | Dubovi, E.J.
A total of 21 calves were inoculated IV with 1 of the following isolates of bovine viral diarrhea virus (BVDV); CD87 (n = 10), NY-1 (n = 3), NADL (n = 5), or were sham-inoculated with virus-free medium (n = 3). Subsequent to inoculation, platelet counts were monitored to detect differences between noncytopathic (CD87, NY-1) and cytopathic (NADL) isolates in their ability to induce thrombocytopenia. Platelet count decrease throughout infection was statistically analyzed by comparing the slope of the line drawn from the count on the day of infection to the lowest count achieved by that calf. Significant difference was observed in the CD87-inoculated calves and in the NY-1-inoculated calves, compared with those of the same control group. Significant difference was not observed in the slope of platelet count decrease between the cytopathic NADL-infected calves and control-group calves. The data indicate that noncytopathic BVDV isolates may more easily induce thrombocytopenia than do cytopathic isolates in immune-naive, immunocompetent calves; acute infection with 1 cytopathic BVDV isolate (NADL) did not induce thrombocytopenia. In addition, although each calf seroconverted, virus was rarely isolated from mononuclear cells obtained from calves with cytopathic infections. At some point after infection, virus was always isolated from each of the calves undergoing noncytopathic infections, and occasionally, transient association of noncytopathic BVDV antigen with platelets was observed during these infections.
Show more [+] Less [-]Characterization of the hemodynamic and metabolic alterations in the large colon of horses during low-flow ischemia and reperfusion
1994
Moore, R.M. | Muir, W.W. | Bertone, A.L. | Beard, W.L.
Effects of low-flow ischemia and reperfusion of the large colon on systemic and colonic hemodynamic and metabolic variables were determined in horses. Twenty-four adult horses were randomly allocated to 3 groups: sham-operated (n = 6), 6 hours of ischemia (n = 9), and 3 hours of ischemia and 3 hours of reperfusion (n = 9). Low-flow ischemia was induced in groups 2 and 3 by reducing colonic arterial blood flow to 20% of baseline. Heart rate, arterial blood pressures, cardiac index, pulmonary artery pressure, right atrial pressure, and colonic blood flow were monitored. Arterial, mixed-venous, and colonic venous blood gas and oximetry analyses; PCV; and blood lactate and pyruvate and plasma total protein concentrations were measured. Data were recorded, and blood samples were collected at baseline and at 30-minute intervals for 6 hours; additionally, data were collected at 185, 190, and 195 minutes (corresponding to 5, 10, and 15 minutes of reperfusion in group-3 horses). There were no differences among groups at baseline or across time for any systemic hemodynamic or metabolic variable. Colonic blood flow did not change across time in group-1 horses. Colonic blood flow significantly (P < 0.05) decreased to 20% of baseline at induction of ischemia in horses of groups 2 and 3 and remained significantly decreased throughout the ischemic period in horses of groups 2 (6 hours) and 3 (3 hours). Colonic blood flow significantly (P < 0.05) increased above baseline by 5 minutes of reperfusion in group-3 horses. Colonic oxygen delivery and oxygen consumption, and colonic venous pH, PO2, percentage saturation of hemoglobin, and oxygen content were significantly (P < 0.05) decreased within 30 minutes after induction of ischemia in horses of groups 2 and 3; colonic venous PCO2, colonic oxygen extraction ratio, and lactate and pyruvate concentrations were significantly (P < 0.05) increased by 30 minutes of ischemia. These alterations continued throughout ischemia, but within 5 minutes of reperfusion in group-3 horses, these variables either returned to baseline (pH, PCO2, lactate, pyruvate), significantly (P < 0.05) increased above baseline (PO2, oxygen content, % saturation of hemoglobin), or significantly (P < 0.05) decreased below baseline (colonic oxygen extraction ratio). Colonic oxygen consumption remained decreased during reperfusion in group-3 horses. Colonic mucosal ischemia-reperfusion injury observed in this model of ischemia was associated with local colonic hemodynamic and metabolic alterations in the presence of systemic hemodynamic and metabolic stability. Reactive hyperemia was observed at restoration of colonic blood flow in group-3 horses and persisted during reperfusion. Colonic venous metabolic alterations were corrected at reperfusion, indicating adaptation of the colon to the return of blood flow and oxygen delivery with resultant decrease in anaerobic metabolism. The early alterations in these variables may simply represent a washout of metabolic by-products.
Show more [+] Less [-]Acute effect of hydralazine administration on pulmonary artery hemodynamics in dogs with chronic heartworm disease
1994
Atkins, C.E. | Keene, B.W. | McGuirk, S.M. | Satō, Tōru
In an effort to better understand the role of vasodilators in the management of pulmonary hypertension associated with chronic heartworm disease (HWD), pulmonary hemodynamic measurements were obtained from 7 experimentally infected, anesthetized dogs before and after hydralazine administration (mean dose, 1.96 mg/kg of body weight). Five dogs were maintained on room air, while 2 were maintained on 100% oxygen during the hydralazine study. The hemodynamic effect of hydralazine in dogs with HWD was evaluated, using heart rate, cardiac index, mean pulmonary artery pressure, mean arterial pressure, total pulmonary resistance, total systemic resistance, total systemic resistance/total pulmonary resistance, left ventricular dP/dt(max), left ventricular end diastolic pressure, and left and right ventricular double products ([mean arterial pressure X heart rate] and [mean pulmonary artery pressure X heart rate], respectively). Responders were defined as those in which total pulmonary resistance decreased greater than or equal to 20% without an increase in mean pulmonary arterial pressure and in which heart rate increase was less than or equal to 10%. Comparison was also made between maximal hemodynamic effect of hydralazine with that after 100% oxygen administration for 15 minutes to previously normoxemic dogs (n = 5). Significance was determined if P < 0.05, using the paired t-test. Hydralazine induced significant reductions in mean pulmonary and systemic arterial pressures and total pulmonary resistance, with no significant change in heart rate, cardiac index, total systemic resistance, left ventricular dP/dt(max), left ventricular end diastolic pressure, or right and left ventricular double products. Four (57%) of the 7 dogs studied were considered responders. Pretreatment cardiac index, mean pulmonary artery pressure, and total pulmonary resistance did not allow differentiation of responders from nonresponders. However, pretreatment right ventricular end diastolic pressure was significantly less in responders than in nonresponders. Two dogs sustained hypotension after hydralazine administration, but no dogs had significant tachycardia. In dogs with experimentally induced HWD, treatment with hydralazine had significantly greater effect on cardiac index and mean pulmonary and systemic arterial pressures and resistance than did administration of 100% oxygen. These data indicate that further study of vasodilators for treatment of HWD-induced pulmonary hypertension may be warranted.
Show more [+] Less [-]Genus-specific detection of salmonellae in equine feces by use of the polymerase chain reaction
1994
Cohen, N.D. | Neibergs, H.L. | Wallis, D.E. | Simpson, R.B. | McGruder, E.D. | Hargis, B.M.
Members of the genus Salmonella were identified in feces from horses, using the polymerase chain reaction (PCR) and genus-specific olignucleotide primers. Feces from healthy horses were determined to be culture-negative for Salmonella spp. Fecal samples were inoculated with known numbers of colony-forming units (CFU) of S anatum, S derby, S enteritidis, S heidelberg, S newport, and S typhimurium. The DNA was extracted from fecal samples and amplified by PCP, using genus-specific primers. Sensitivity of the assay extended to 10(3) CFU of Salmonella sp/g of feces, sensitivity of microbiologic culture with enrichment extended to 10(2) CFU of Salmonella sp/g of feces. Feces that were not inoculated with Salmonella spp were negative by the PCR. Detection of salmonellae in feces was possible, using the PCR, within 10 to 12 hours from the time of submission of samples.
Show more [+] Less [-]Histologic and ultrastructural changes after large-colon torsion, with and without use of a specific platelet-activating factor antagonist (WEB 2086), in ponies
1994
Wilson, D.V. | Patterson, J.S. | Stick, J.A. | Provost, P.J.
The role of platelet-activating factor (PAF) in mediating the colonic damage that develops after large-colon torsion was studied in 14 ponies. Morphologic changes in areas of the ascending colon and selected abdominal and thoracic viscera after 1 hour of large-colon torsion and 3 to 5 hours of reperfusion were determined, as well as the protective effects of systemic administration of a specific PAF antagonist (WEB 2086). Ponies were selected then allocated at random and in equal numbers to 2 groups that received 1 of 2 treatments prior to induction of large-colon torsion: group 1 - control (saline solution), and group 2 WEB 2086 (3 mg/kg of body weight loading dose and 3 mg/kg/h for the remainder of the study). In each pony, full-thickness tissue specimens from the gastrointestinal tract-cecum, pelvic flexure, left and right ventral colon, and right dorsal colon - heart, left lung, liver, left adrenal gland, spleen, and right kidney were collected and histologically evaluated. Edema, mucosal necrosis, and neutrophil infiltration in colonic sections were graded from 0 (normal) to 3 (most severe changes). Sections of liver and lung from 3 ponies in each group, and colon from 1 pony in each group, also were examined by transmission electron microscopy to determine the presence of ultrastructural alterations. Ischemia and reperfusion induced marked changes in all sections of colon in all ponies: moderate to severe submucosal edema, moderate necrosis of the superficial epithelium and lamina propria, and necrosis of the mucosal crypt epithelium. Extra- vascular neutrophil accumulation was evident in all sections of colon and cecum, but not in other tissues. Ultrastructural lesions were not present in hepato- cytes or pneumocytes, or in the endothelial cells of liver, lung, and colon. Bacteria were observed by electron microscopy in 5% of hepatic sinusoids. Administration of a specific PAF antagonist, WEB 2086, failed to reduce severity of the observed lesions, indicating that it was not cytoprotective at the dosage used in this model of ischemia-reperfusion injury.
Show more [+] Less [-]Characterization of an Actinobacillus pleuropneumoniae seeder pig challenge-exposure model
1994
Lechtenberg, K.F. | Shryock, T.R. | Moore, G.
Five strains of Actinobacillus pleuropneumoniae serotype 1 were used to intranasally infect 5 groups of pigs. Using each bacterial strain, infected pigs (termed seeder pigs) were commingled for 48 hours with 5 groups of noninfected test pigs, then were removed. Seeder and test pigs were maintained in isolation and were observed for 14 days. Seeder pigs had mortality that was threefold greater than that of test pigs (24% vs 8%). Rectal temperature in excess of 40.3 C was achieved for 84% of test pigs and 88% of seeder pigs. Neither of these 2 variables was statistically different between the 2 groups of pigs. Clinical impression scores greater than or equal to 2 (on a 0 to 3 scale) were three-fold (64% vs 20%) greater for seeder than for test pigs (P < 0.05). The total number of bacterial isolations or nonrecoverable isolates was tabulated for test and seeder pigs' lungs at necropsy, irrespective of the amount of lesions. The number of A pleuropneumoniae isolations was not statistically different between test and seeder pig populations. Recovery of Pasteurella multocida or other bacteria was greater from the seeder pigs (P < 0.05), whereas the number of non-recoverable isolates was greater from test pigs than from seeder pigs (P < 0.05). Assessment of lung lesions at necropsy by either visual estimation or on a weight basis were in agreement. Fewer test pigs had lung lesions in excess of 5% of total lung volume than did seeder pigs (40% vs 84%) and, according to the odds ratio estimation, seeder pigs were 7 times more likely than test pigs to have such lesions. These results indicate a predictable, moderate intensity, natural exposure model for use in the study of Actinobacillus pleuropneumoniae-induced pneumonia. The seeder pig model appears to mimic field infection in development of clinical illness, febrile response, lung lesions, mortality, and low potential for secondary pneumonic bacterial involvement, thus providing a useful tool for preliminary evaluation of anti-infective modalities.
Show more [+] Less [-]Immunomodulatory effects of staphylococcal antigen and antigen-antibody complexes on canine mononuclear and polymorphonuclear leukocytes
1994
DeBoer, D.J.
Staphylococcal antigens and immune complexes (IC) prepared from antigen and hyperimmune canine serum were tested for their effects on certain functions of mononuclear (MN) and polymorphonuclear (PMN) leukocytes (cells) obtained from healthy dogs. The effect on MN cells was studied by determining the ability of antigen or IC to augment or inhibit mitogenesis induced by phytohemagglutinin (PHA). The effect of antigen or IC on PHA cells was studied by measurement of H2O2 production as an indicator of respiratory burst. Neither the antigen nor the IC, when cultured with MN cells, was mitogenic. Coincubation of antigen or IC with MN cells and PHA resulted in a concentration-dependent decrease in mitogenesis. The decreased mitogenesis could not be overcome by addition of excess PHA, and may in part have been related to toxic effects of the antigen or IC on MN cells. When MN cells were instead preincubated with antigen or IC, then washed and stimulated with PHA, there was still a concentration-dependent inhibition of mitogenesis, although toxicity to the cells was not observed. Low concentrations of staphylococcal antigen or IC stimulated slight H2O2 production by PHA cells. When PHA cells were coincubated with IC and another stimulus (opsonized zymosan or phorbol myristate acetate), IC appeared to augment phorbol myristate acetate-, but not zymosan-induced stimulation. These results suggest that staphylococcal antigens, either alone or complexed with antibody, have the ability to stimulate PMN cells and inhibit MN cell function. Such actions may have a role in the pathogenesis of recurrent staphylococcal infection in canine patients.
Show more [+] Less [-]Effects of subject stance time and velocity on ground reaction forces in clinically normal Greyhounds at the walk
1994
Roush, J.K. | McLaughlin, R.M. Jr
Force plate gait analysis was used to study the effects of subject stance time and velocity on ground reaction forces in 5 adult Greyhounds at the walk. Data from 146 valid trials were obtained. Stance time and velocity were linearly related, and stance time had a strong, negative correlation with velocity k = -0.72 for the forelimbs, r = -0.56 for the hind limbs). Stance time correlated more closely with changes in peak vertical force and impulse than did velocity. Stance time and velocity correlated less strongly with braking and propulsion forces and impulses. The trials were divided into 2 distinct velocity ranges (V1 = 0.92 to 1.03 m/s, V2 = 1.06 to 1.17 m/ s), 2 distinct forelimb stance time ranges (FST1 = 0.43 to 0.48 second, FST2 = 0.50 to 0.55 second), and 2 distinct hind limb stance time ranges (HST1 = 0.40 to 0.45 second, HST2 = 0.46 to 0.51 second). Five trials from each dog were included in each range, and the mean values were used to evaluate changes in ground reaction forces between groups. Peak vertical force in the forelimbs decreased significantly (P = 0.048) as FST increased; however, difference was not detected in vertical force between velocity groups. Peak vertical force in the hind limbs decreased significantly (P = 0.001) as HST increased and increased significantly (P = 0.000) as velocity increased. Differences were not observed between groups in forelimb or hind limb braking and propulsive forces. Vertical impulse in the forelimbs and hind limbs decreased as velocity increased and increased as stance time increased. Braking impulse in the forelimbs decreased as velocity increased and increased as FST increased. Braking force in the hind limbs did not change between velocity or stance time groups. Propulsive impulse in the hind limbs decreased as velocity increased and increased as HST increased. Stance time was a sensitive and accurate indicator of subject velocity in clinically normal dogs at the walk and correlated more closely with changes in some ground reaction forces than did velocity measurements. Stance time measurements could be used to normalize trial data within a sampling period and document consistency in velocity during force plate analysis of clinically normal dogs at the walk.
Show more [+] Less [-]Consideration of anatomic and radiographic features of the caudal pouches of the femorotibial joints of horses for the purpose of arthroscopy
1994
Development of an arthroscopic approach to the caudal pouches of the equine stifle has been necessary because cranial approaches do not allow access to articular lesions in the caudal aspect of the joint. Therefore, the anatomy of the caudal region was examined in 52 cadaver limbs by use of gross dissection, x-ray-computed tomography, fluoroscopy, or arthroscopy. Additionally, using arthroscopic techniques developed in equine cadaver limbs, 3 stifles from 2 anesthetized horses were arthroscopically explored. Fluoroscopy was used to verify needle placement for joint injection and filling patterns of each femorotibial joint. The medial femorotibial joint sac (n = 4) held a mean +/- SD 41.67 +/- 5.77 ml of injection fluid, and the lateral femorotibial joint sac (n = 4) held a mean 61.67 +/- 2.89 ml of injection fluid. Vital structures that inadvertently could be damaged during arthroscopy of the caudal pouches of the stifle included the peroneal nerve (located approx 7 cm caudal to the lateral collateral ligament), the popliteal artery and vein (situated directly between the medial and lateral femoral condyles), and the lateral femoral condyle (most often traumatized during arthroscopy). The tendon of the popliteus muscle, which is contiguous with the joint capsule of the caudal pouch of the lateral femorotibial joint, made arthroscopic exploration of this pouch particularly difficult.
Show more [+] Less [-]Quantification of serum total IgE concentration in dogs by use of an enzyme-linked immunosorbent assay containing monoclonal murine anti-canine IgE
1994
Hill, P.B. | DeBoer, D.J.
A method for quantification of serum total IgE concentration in dogs by use of an ELISA containing monoclonal mouse anti-canine IgE was developed. Microtitration plates were coated with monoclonal mouse anti-canine IgE. Test sera and reference serum dilutions were added, followed by biotinylated monoclonal mouse anti-canine IgE. Avidin-alkaline phosphatase conjugate was added, and color development was measured spectrophotometrically, using a microtitration plate reader. Quantitative results were obtained by assigning to a reference serum a value of 100 IgE units/ml. Absorbance values of unknown samples were converted into IgE units by comparison with a standard curve generated by measurement of reference serum dilutions. Intra- and interassay coefficients of variation were 5 and 7%, respectively, and assay sensitivity was 1 U/ml. The assay was used to establish a normal range for total IgE concentrations in 30 healthy dogs. Total IgE concentration in healthy dogs followed a skewed distribution and ranged from < 1 to 91.2 U/ml, with a geometric mean value of 7.1 U/ml. The IgE concentration was remarkably stable in serum samples subjected to 25 freeze/ thaw cycles or incubation at approximately 25 C (room temperature) for up to 10 days. Comparison of total IgE concentrations in 23 serum samples assayed by use of double-overlay radial immunodiffusion and ELISA yielded correlation coefficient of 0.94. Comparison of the reference serum standard curve with serial dilutions of a purified IgE solution of known concentration yielded a range of values for the IgE unit of 0.7 to 2.0 micrograms.
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